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One Source. One Search. One Solution. 

IN VITRO MICROBIOLOGICAL MUTAGENICITY 

STUDIES OF FOUR CIBA·GEIGY CORPORATION 

COMPOUNDS (FINAL REPORT) WITH TEST DATA 

AND COVER LETTER 

08 MAR 1990 

U.S. Department of Commerce 

National Technical Information Service 
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/'i~"":\ U.S DEPARTMENT OF COMMERCE 

. 1Jj'' . Technology Administration 

\ ·r / National Technical Information Service 

..._., Springfield, VA 22161 
_C_9PING FORM FOR SRC INDEXING, REVISED 10'115185 

IN VITRO MICROBIOLOGICAL MUTAGENICITY STIJDIES OF FOUR 

CI-...sEISV CORPORATICJt.! COMPOUNDS (FINAL REPORT) WITH TEST 

DATA AND COVER LETTER 

He:XABROMOCVCLODODECANE (3194-55-1!;) 

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GreafLakes-. 

Chemical Corpitratio~· 

March 8, 1990 

Document Processing Center (TS-790) 

Room L-100, Office of Toxic Substances 

Environmental Protection Agency 

401 M street, sw 

Washington, DC 20460 

Attn: 8 (d) Health & Safety Reporting,, 

(Notification/Reporting) 

Dear Sir: 

Contains . N'"· u ·· ca~ j 

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In response to the TSCA Section 8 {d) rule' published in the December 

12, 1989 Federal Register, the enclosed information is being sub- , mitted on the followin~ substances: 

The 

the 

2,4,6-Tribrornophenol 

'Tetrabrornophthalic anhydride 

Hexabromocyclododecane 

1,2-Bis(tribrornophenoA~)ethane 

Decabromodiphenyl ether 

Pentabromodiphenyl ether 

Octabromodiphenyl ether 

studies on the first ,four compounds 

following manufacturing facility: 

(., 

Great Lakes Chemical' corporation 

Route 7, Box 529 

El Dorado, AR 71730 

cAs Number 

118-79-6 

632-79-1 

3194-55-6 

37853-59-1 

1163-19-5 

32534-81-9 

32536-52-0~-

; :. 

are submitted on behalf of 

The studies on the last three cornpou."lds are su~mitted on behalf of 

the fcil:J.owing manufacturing facility: ' •, 

Great Lakes Chemical Corporation 

'' Highway 15 South, P.O.Box 1878 · El Dorado, AR 71731-1878 

' 
r;. 

" There are no known additives to these compounds. Pentabromodiphenyl ether consists of hexa-, penta-·, tetra-bromodiphenyl ether. 

Octabromodi.phenyl ether consists of nona-, octa-., hepta"'-, and hexa- " 

bromodi.phenyl ether. ·we are not aware of any ongoing studies with 

any of the listed compounds. If you have any questions concerning this submission, please call me at (317) 497-639l.: ·--. 

Sincerely, 

J).L-tt"-d/~ /..{<f~ 

Dennis L. McFadden, Ph. D. 

Product Safety Coordinator ,. 

DLM:sb:OJOS90 

Enclosures 

;.-::. <.: 

·' . 
II 

TEST DATA 

2,4,6-Tribromophenol 

Acute toxicity studies in rats and rabbits 

Primary skin i=itation study in albino rabbits 

Eye! i=itation study in albino rabbits 

Acute inhalation toxicity in the albino rat 

Acute dermal toxicity in male and female albino rabbits 

'Acute oral toxicity (LD50) study in albino rats 

Dermal sensitization study in the albino guinea pig Bioaccumulation study in fat tissue with 2,4,6-tribromophenol 48-Hour dynamic aquatic toxicity study with 2,4,6-tribromo- phenol in Daphnia Four-~ay static aquatic toxicity studies with 2, 4 , 6-tribromo- .. • 

phenol in rainbow trout and bluegills. Screening test for microbiological inhibition with 2,4,6- tribromophenol Acute dust inhalation toxicity study with 2,4,6-tribromo- phenol, lot number 3106 in rats · 28-Day subacute dermal toxicity study with 2,4,6-tribromo- phenol in albino rabbits (10/27/77) 28-Day subacute dermal toxicity study wth 2,4,6-tribromo- phenol in albino rabbits (12/12/77) · Pilot teratology study in rats 

Mutagenicity evaluation of 2,4,6-trib:i:oroophenol, lot number 

3287, in the Ames Salmonella/Microsome Plate Test 

Acute oral toxicity (LD5J study in rats ' =· 

The bioaccumulation of 2,4,6-tribromophenol in bluegill sunfish 

2,4,6-Tribromophenol biodegradation (9/30/75) 2,4,6-Tribromophenol (TBP) biodegradation (6/1/76) Pharmacokinetics study of 2,4,6-tribromophenol in rats 

Photolysis of 2,4,6-tribromophenol ·partition coefficient of several flame retardants and 

industrial chemicals 

Water solubility of several flame retardants and-\ndustrial chemicals -- Photolysis of 2,4,6-tribromophenol in aqueous solution 

,, 

. ~·. 

' 
Tetrabromophthalic Anhydride 

Acute oral study 

Acute dermal study 

A.cute toxicity studies of tetrabromophthalic anhydride 

Primary skin irri.tation study in albino rabbits 

Eye irritat:i.'On study in albino rabbits 

Acute inhalation toxicity in t;he albino rat 

Dermal sensitization study in the albino guinea pig 

Acute inhalation toxicity in the albino rat after'pyrolysis 

(3/12/75) r;?' 

Acute inhalation toxicity in the albino rat· after pyrolysis 

(3/26/75) 

28-Day dermal toxicity study in rabbits 

21-Day inhalation toxicity in rats 

Human repeated insult patch test with Firemaster PHT4 

Mutagenicity_evaluation of 859-74-4 

Pilot teratology study in ra't:s ' ' 

The acute toxicity of Firemaster PHT4 to the water flea, . Daphnia magna Straus 

The acute tox~city of Fi.,emaster PHT4, lot number 6332-B 

to the rainbow trout Salmo gairdne:o:i Richardson 

The acute toxicity of Fire~aster PHT4, lot number 6332-B 

to the bluegill sunfish Lepomis macrohirus Rafinesque 

Acute oral toxicity study in mice 

Comparative inhalation toxicity tests '-~ 

Evaluation of the inhalation toxicity of combustion products 

of selected polymers II 

The. bioaccumulation of tetrabr·omophthalic; anhydride in the 

bluegill sunfish '' 

\; Persistence of PHT-4 in soil at 1 and 10 p.gjg 

Photolysis of PHT-4 

.Effects of PHT-4 on soil microflora.at 1 and 10 p.g/g 

''Pr.armacokinetics of PHT-·4 in r-ats 

Partition coefficient of several flame retardants and 

industrial chenicals 

Water solubility of several flame retardants and industrial 

chemicals 

,, 

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Hexabromocyclododecane 

/.Hexabromocyclododecane: 28-day feeding trials with rats and 

90-day feeding trials witll rats 

VIn vitro microbiological mutgenicity studies of four CibaGeigy Corporation compounds Pilot cataractogenic study in chicks 

cataractogenic study in chicks 

Mutagenicity evaluation of 421-32B 

Acute toxicity studies in rabbits ad rats 

Mutagenicity of two lots of FM-100, lot ~3 and residue of 

lot 3322 in the absence and presence of metabolic 

activation ,; .. , ·· 

The acute toxicity of HBCD lot number 990-17 tq the bluegi~l sunfish Lepomis macroh'irus Rafinesque 

Partition coefficient of dicamba, endrin, VEL 3510 and' 

several industrial chemicals and flame retardants 

Water solubility of several industrial chemicals, flame 

retardants and a herbicide VEL 3510 

Hydrolysis of Firemaster 100 

HBCD - biodegradability study 

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" 

" 
,, 

,, 

1,2-BisCtribromophenoxylethane 

l.i 

-water solubility of several flame retardants and industrial chemicals 

-Photolysis of Firemaster 680 

-Pharmcokinetic study of FM-680 -Rate of hydrolysis studies 

-Partition coeffi.cient of MC-680, chlorendic anhydride and chlorendic acid 

-28-Day rat feeding study -Acute oral toxicity study in the albino rat -Acute dermal LD50 (l level) -Acute inhalation toxicity in the albino rat -Acute oral toxicity study in beagle dogs --il.cute toxicity (TL.,; 0 ) study in the bluegill -Acute toxicity (LD50 ) study in the rainbow trout -Acute heated vap.or inhalation toxicity study in rats -Modified Draize niul tiple .. insult test in male humans -28-Day dermal toxicity study in rabbits ~21-Day inhalation toxicity in rats -Acute inhalation toxicity in the albino rat after pyrolysis (ABS resin/FM-680/Sb2

0.3) Acute inhalation toxicity in the albknO rat after pyrolysis (~BS ;:esin/~b ) ._14-Day range fnd~ng study Ln rats 

'28-Day toxicity study in rats 'Biodegradation study with' 14C-tagged MC-680 -Biodegradability study"with 14c-tagged MC-.680 " "Primary skin irr·itation test with mother ·liquor 680 

process, code no. 7-29-76-B in albino rabbits -skin sensitization test with mother liquor 680 process in albino guinea pigs """'90-Day subacute oral toxicity study with Firemaster 680 in albino rats (Jan. 6,: 1977) .:• 

'90-Day subacute oral toxicity study with Firemaster 680 

final report 

1."; 

in albino rats (Mar. 10, 1977') 'Mutagenicity evaluation of 859-74-5 --Test report (method for testing the 

of chemical substances in fish in fish) · 

degree of accumulation 

boay) (Bioaccuniulatiofi 

,,, "'Acute inhalation toxicity study in the albino rat after pyrolysis (ABS resin/Sb2

0;Vcontaining 680) Pilot teratology study in rats · 'Teratology study in rats ' "Tissue residue accumulation/depletion study with Firemaster 680 in albino rats · ,, 

,, 
Decabromodiphenvl ether 

\Acute toxicity studies in rats and rabbits 

\Mutagenicity evaluation of componrid 364-5A 

; .'-28-Day. toxicity study. -in.,rats·..:.·.(-5/-12/-7~).,.'=·'==--=;.-'----"'· 

'28-Day toxicity study in rats (6/24/76) '28-Day toxicity study in rats (3/2/77) ~Mutagenicity evaluation of compound 277-10 

Pentabromodiphenyl oxide 

-r!utagenicity evaluation of compound 345-76A final report, 1

, 'The acute oral toxicity of pentabromodiphenyl ether to rats· 

\Acute toxicity studies in rats and rabbits 

\28-Day toxicity study in rats 

'The bioaccumulation of compound S-512 by carp "'Jo-oay dietary study in rats with pentabromodipl'i:enyl oxide 

including recovery periods of 6, 12, and 24 weeks ~ 

'90-Day dietary study irrrats with pentabromodiphenyl oxide (DE'-71) ,, 

Octabromodiphenyl oxide a 

J The bioaccumulation of compound S-511. by carp ..-1\cute toxicity studies in rats and rabbits 

v1subacute inh.alation toxicity study in rats ~Mutagenicity evaluation of compound 34S~79A final report ., Jrn vitro sister chromatid exchange in Chinese hamster• ovary cells with octabromodiphenyl ether (DE-79) Junscheduled DNA synthesis assay, compound DE-79 ~28-0ay toxicity study in rats 

'13-Week feeding study in rats 

'A range-finding teratology study in rats with DE-79 

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000b3b728hl 

, '23'.b-40Dbt:)C::, &-f:.;}. .October ll; 1976 

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Final Report 

ltl Vl'rnO :.l!CROBIOI.OGICAL MUTAGENICITI(, STUDIES 

'OF 'ifcii:iit cuiA-GEI Gi ;CORPORATION cm.IPOU.NDS .:: 

• 

Prepared :for: 

. f(.e '0-.br'dl~t/jc14.<~d~ 

.·' cAr. 1:/1=. 3'trt--sr-(, ' CIDA~GEIGY CORPORATION 

ArdslP.y, New York 1.0502 

A ttent19n: Dr. Martin G. Bernstein 

Submitted by: · 

Vincent ·F. SJ.nunon, :.tnaager 

Microbid.l Genetics Program 

Denis c. Poole 

M{c:robiologist 

SRI Project LSC-5702 

Approved: ,, 

if.!~ w. /frv~ Gordoh ·w. Newell, Director 

Department of 'I'oxicolO'gy 

vu. & . w.- A. Skinner, Executive DircCto:i= 

Life Sciences Division 

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sm.t:.tr\RY . 

sni tes:tcd :four CIBA-G:E:IGY CorP,,or;~tion co::~poundn to evaluate 

,their potential "for DUt:tceneSis in five S"tr:11ns of Salmonella 

typhirnuriurn--TAl5351 TA15371 TA15381 

wu.-5 pe(t".formed in the presence and i.n 

TA981 and TAlCO, Each ~ss~y 

' 0 .;:1 c. the absence of n~metnbolic 

system. Compound'42l-32-A was very 

.}' ' d) 

mut:.genic "1n strains 

,, 

nc:t1vat1on 

TA1'!;35 and TAlCO when the rat-liver~homogenate metabolic oct1 v~ t.1an o 

' system ~fS added to the aas~y. 7he three other compounds, 421-~2-B1 

421-:32-c, and !.!A-261 were 

nssays 1n t~B presence on 

system. 

.. " .:: 

not mutage~c in any o: the bacteria~ 

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in the absence ot a metabolic act1.yat1on 

il2~-32B=(Hexabromocyc~ododecane 

No. 760403~8) o 

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SRI examined :four CIBA-GEIGY Corpor.at:1.on compounds for r.nit::'.l'-

gen1c1 ty by .!,:: ~ l':licro01.ologicOJ.l as~.ays w1 th 1'1.ve strains ?:r 

S3lr.tonell:J typhimur1um. An Kroclor 1251-stimula t~·d~ rrit-liverhomogenate metabolic activation sy~tem was :1.ncluded in ~he assay 

.. 0 

procedure • 

incapable 

to provide metabo~ic steps that the;bac~Cria e1ihe:r: are ·.f:: 

o:f condtict:1ng or do not Carry out under the a~~3y cond.1 t!Ons.;.:-

The P.urpose of this study was to determi.ne whether thtj compound~ 

elicited a mutngenic response ;tn miCroorganism~. 

The assay p;rfOcedure w1th S. typh!:muriurn has been proven t.o be 

85 to 90% accurate 1n detecting carcinogens :a~~ mutagens," and it b.:ls .. • ? • ·-

·.about the saq1e accuracy in -1dentif;Yi.ng chemicOJ.ls that are- nOt Carci.no-· 

~~nic .. -

1 How7ver, because the test \y~1:em .is not 100~ accura te1 

neither a 'pos1 ti ve nor a negative re_~ponse prC?ves th::1 t :l chem1.cal 

c 1s h~zordous or nonh;Iz:.trdous to rnan. 

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r.1ETitODS 

Salmonelln typhimurlum Strfiins T•'l535, T,'\1537, TJ'l538, 

TA!JS~ :md T;\100 .. 

The Salmonelln typhirn.urium st:ra1ns used at SRI :n·e nll histidine 

nu otrophs by virt~e ~:f mutationS in the histid1"ee operon. When" these 

hiStidlne-depfmdent ce~ls !.~:re grown on a rn1nimnl media petl·i plnte 

deficient in h:1stid1ne', 

• + independence (his ) are 

onl;r those cells that revert to hl.Stid .. ine 0 ' . 

able to grow (mutants). The spontaneous 

mut.<l'"tion !req~ency Or each strain 1s rel:ltively constant, but when 

a. mutagen is added ~0 the :t{;':Jr~'the mutation frequency is increased 

3 to ;J.OO times. 

\'le obtained our!· '~'typhirnul·ium strnins from Dr .. , Eruce Ames of the 

University of Cali:forn~a nt D~rkeley. "::"s In :Jddi_tion to having ~=tutations in the histidine operon, all the i.rid.icntor strains hnvc mutations 

in the lipopolysacch::trlde coat (r.fa -j :md deletions that cover a gene 

involved in the repair o! uv d:un:1ge (~D-L The :rfa- mutation makes •.i --

the ntrains more perrneabtc. to l:xrg:e r:;olccu"les, thereby increasing 

their ~ensi ti\.'1 ty to these molccti'lcs. tJ 

The uvrB- mutation decreases 

repair o£ some types o! Chemically darn3ged D~A and thereby enh~nces 

sens~'ti Vi ty to some r.tut.;;~.genic chectic:llS·. Strairi TJ'l535 is reverted 

. + ' 

to hiStidine prototrophy (his ) by foiany r.mtagcns t.h:Jt ,cause J>ase-pnir -:; ' 

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substitutions. 7~100 iS derived from ·TA1535 by the introduction .:: 

the n ·'fnc~or plas~~d ptu.tlOl. G The i:~troduction Of ~hiS plasmid, 

0 ·-~~-·= ~ 

··hi h 'le c: a. ... 

~.f t.) m·~~ ,~ ~ t \1~ '.1 c ...... ~ "!:.. 9: ~ c:: ;') ..... :'t' c':'nfers ampicillin r-esistance to the stt·ai:', ¥':rcatly enhnncris tJ1e ~1 

~ens1tiv1.ty of the strnin to some b~se-~~i'~-.-~ubsti1:u1:ion mu~::agens.. "~~·~8-~ .. ~ 

\'le hz.ve shown ihat mutagens such _as b~nzYi chlol?dc omd 2-{2-furyi.)_-

3-(5-nitro-2-furyl) ocrylnmide (known'os AF2) con: be detected in plnte 

assays by TAlOO but not b~· "f}\1535. The pre:sence:.·~f this plasmip alsO 

makes strain T.\100 sensitive to ·same frame~hift ~~tngens--e.g·., 0ICR-191, . : .. 

benzo(o)pyrene, alfatoxtn EF and 7,12-dimethy1benz(a)nnthiitcene. ~ .: ~ '·~~~ ' ~ 

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St:r~1ns TJ\1537 ond 'TJ\1538 ::arc reve:rtod by m~ny frnmeshift mutogcns. 

T/1 1537 is more sens~tive th~n TJ\1538 to muto:ltion by some acrldi.ncs and 

ben::anthracens.7 but the difference is quantit:Jtive r.nther th::m qualita- .. 

tive. TA9S is derived from T/\1.538 by the addition of the plasmid pi-::.MlOl, 

which makes this str.nin more sensitive to ~orne muta~;cns. . ~ 

All the indicator strains are stored ot -80°Cn For each experiment, ::m 1i1oculum :from frozen stock cultures is grown ov:ern1ght at 

37°C in a nutrient broth consisting of 1% -tryptone :Jnd 0.5~ yeast 

extract. After stationary overnight growth, the cultures are shaken 

for 3 to 4 hours to ensure optimal gr~wtb. Each culture is checked 

for sensitivity to crystal violet. The presence of the rfa- mutation 

makes !i'the indicator strains sensi.tive to this dyr, whereas the parent 

+ .. . strain, !!! , is not sensitive to the dye. Howeve~, the mutation is 

• ' + ' rcversi.ble, leading to the accumulation o:t: rfa cells i.n the culture. 

!) ·-- • ' 

Therefore,, the cells must be tested routinely to ensure thei-r sensitivity to c:r·yst:al v:1.olet. Eoch culture also is tested by specific 

mutagens known to revert each test strain ( pos1 ti.ve controls). 

AsSnys in Pgnr 

To a sterile 13 x 100 l{tl'n test tube placed i.n a 43°C heating 

block., we add in the following order: 

(l} 

(2) 

q 

2 ml of 0.6% agar o 

0 .. 05 ml o! indicator organ.isms 

(3) 0.5 ml of metabolic activation mixture (optional) 

(4) Up to 50 IJ.l of·' a solution of the test chemical. 

For negative-controls, we use'steps (1), (2}, and (3) (optional) 

and 50 lJ.l o! the solvent Used for the test chemical. 

This mixture is sti.rred gently and then" poured onto r.tinimal agar 

plntes~t After tt1e soft agar has set, the plates are incubated at 

q. 6% agar contains 0.05 rn.\1 histidine and 0.05 m.\l biotin. 

1 Minimal agar plates consist of 15 :-g of agar, 20 g of glucose, 0.2 g 

oi MgS04 '7 .H?O, 2 g o! citric ,acid monohyd:atc, 10 g of K

2

IIPo

4

, and 3.~ g of NafiNH4

Po4 ·H

2

o per liter. 

4 

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The + nur.1ber o! his tJ rcvert:an ts is coun·ted :md recorded .. 

Some of the revert::mi.s ;:arc routinely tested to confirr.t that they arc 

hts+, require b1ot1n1 ::md nre .sensitive t~ cryst::tl violet (rf:a-) • .. , . -- Mct:~bolic .ACti va tion--il rocl\J~" 1254-Stimul:::J ted r.te;:nboli c ~ct1 \."a t1on 

System 

Some carcinogenic mutagens (e .. g. 1 d1rnethyln1tros::am1ne) are 1nnct1ve 

unlc:S~ they nre converted to their ~ct:i.ve fort:'i by betn'g metilbo11zed. 

ftmes has described the rnet:::~bol1c nct1vat1on Systems we use.G1 7 Jldult 

male rats (250 to 300 g) are given a single 500 mg/kg intraperitoneal 

injcc'tion of a polychlorinated biphenyl (J'roclor 1254).7 Four days_ 

after the 1nj;ct1on, the animals' :food is z·emoved.. On the fifth cby, 

the rats are killed and the 11ver homogenate prepared .. 

The livers are removed asept1C4!lly and placed in prcwe1ghed 

sterile glass beakers. 'l'he org:n.n weight is determined, :m~ ::all subsequent operntions to the metabolic activati?n step a:x·e conducted 1n an 

i:_-ce b3th.. The orr;.:Jn 1s w::1shed in ::10 equal volume of' cold, sterile 

0 .. 15 M KCl (lml/g of wet or:;:m)., minced with sterile surbiCill scissors .. 

in three volumes of 0.15 K~l, and ho~ogenized with a Potter-Elvehje~. 

apparatus. The hornogenat~·is centr1~uged fer 10 minutes ~t 9000 x~1 

and the supernatant 1s removed and stored 'in liquid nitrogen. 

The met<lbolic acti\•ation system f'or e::ach experiment consists 

of (for 10 ml): ('! 

1) 1 .. 0 ml• postmitochrondrial sttpernat;J.n!= 

2} 0.2 ml J.!gC12 (0.4 lot) l(Cl (1.65 :.t) 

3} 0.05 ml t;lucose-6-phosphote ( 1 M) 

4} 

5) 

0.4 ml NP.DP. (0.1 lot) 

5.0 ml sodium phosphnte (0.2 M pH 7.4} 

5 . ' 
. -.. : .. 

· Jtl-:SUJ:l'S .1\ND DISCUSSION" 

ImmCdin tcly bciol"c '1scJ c~ch compOund aiid the po::o:l tivc controls 

vterc <ll:.sol\·cd in dimethyi'sulioxidc (D~ISO) so that 50 Ill o! the 

solvent contnlncd the dcsirCct dose of chcr.~.1col. The negative control 

,, wns tl'C:l'l:cd wi lh 50 J.!.l of D.\150. 

fhc rcsul ts pres en ted in "fable 1 nrc the nvcr;:q;c of at le4lst 

two c_::tpcr.lmcn t:s conducted on scx1arn tc dnys. .An cxp~rimcn t consists 

of :.::;soJys with the lcs t compound nnd with posi t1 vc and n ega tl vc 

controls. Eac~ compound is tested first over a wlde rnf1gc of 

.,doses on nll str01ins in the presence ond in the absence of li vcr 

homoccnate for datcrmi.nation of its toxicity ;:md/or mutar:cnicity .. 

li' the compound .c.Jocs not nppcar to be mut::~gcnic or tox~c, it is 

" ·retested for confirmation of this result. 

Compounds 421-32-B_,. 421:--32-c, nnd MA-26 w&rc not mutagenic or 

to:x:ic.. However, the prcliminnry c:x:pC~imci1t wi.th compound -121-32-A 

indicated that it was mut::~cenic on st.r-.ains_ "''Al535 and TAlbO in the 

pr~sence o:f the liver homor:cnate. Two additional experiments were 

pcr.formcd to confirm that 421-~2-A i.s mutnacnic.. Table l presents 

the nvcrage oi these two experiments .. 

Compound 421-32-A was verY mutag-enic when tested Wi.th ratliver-homogenate activa~i.on system, but it was not rnutngcn~c in 

the absence of ·the activation system. The 'compound was more 

mut<Jgcnlc in assays with TAlOO than with TA1.535. Fail~·ly linear 

dose responses were obtained in both Of these strains up to a 

dose of 100 l'g-/plate.. Compound ~21-32-A was wenl<ly mutor;enic 

in TA98 1 .tn "the presence o! the actiVation system.. St:rain T.,\1535 1.s 

reverted· to histidine indcpenedence by mutagens that c~u;e base-pai.r 

substituti:on mut::~tions 1 whereas- strai.n TAlOO is reverted by bothfrnmeshift nnU base-pair substitution mutagens. .-

6 
•. 

• 

.· .... 

- r·•, ,; 

CC:uupc.•tl1at1 ~::n:~J:.!-;\ J.::; n :; ll'OI~l!' mut:~~c.n :t~ltl 1::;: ·COIIl,ilill"OJIJlc J•clot'l.l vo 

lo m11lnscn1c nct:!vlty :.md ns.:;ny cO'ndit:ionsi (i.c. 1 it in ruutact!nic 1~: ' -: :i I,' I • • 

s t.l>n1ns <r· "li\1535 mal "J'J\.100 only when the li vor hoint)gcnil tc is pr<.:~cn t) 

to tris(21 31 -<ll:bromopropyl) phospl1a ta1 ~nether haior;cnatcd al1ph~tc 

phoscphate we h~·ve tested. 

' 

{j 

·)' 

•·· 

,c 

:J 

.. 6 a. 
"' '};,' 

.,;: > 

" :,-,. 

CompoUnd 

lfecntivc control 

Postt1\•e control 

.;-PrOpiol:Jc tono 

AF-2 

9-JlD1noncr1d1no 

2-An,hram1na 

Cl n.I-GEIOY 421-32-A 

Meto.bollc 

Activity 

+ 

+ 

+ 

+ + 

+ 

+ 

+ 

+ 

+ 

,•.,; .. 

' ... 

() 

'l'nble l 

!!!. AS&iYS WITU 'riPHIMURIUM 

,, 

pr. of Compound lidded per Plato /lvcroge·Nurnbor of Uistid.ine-Posttive Revertants por Plau TJ\1537 TA153d 

. 27 9 28 27 165 

19 H - 34 37 163 

50 712 - 782 0,05 

100 312 838 

20 ·48 63 20 " 577 1380 

1 11 10 13 27 170 

s 

10 

25 

8 2 .9 27 ·- 1GS 

ll ,;::9 ":;; 7 11 N 159 

10 1 G 27 1'10 50 

100 

500 

1000 

10 8 ll 33 1G5 

u 8 p " 3·1 152 21 4 11 25 177 

3G 11 10 •27 202 

1 10 15 27 2·1 

5 S2 7 IS 3G - J78 

10 177 11 12 41 .5'21 

25 41ol 12 11 35 . 8·1-1 

100 

sao 

1000 

7!H 

0 

IS 13 5-I l3.:l!l 

1215 19 19 70 2GOG 

2330 28 19 95 1500 

2•JGO ?/:. 29 22 101 <1200 

.... 

•· ·" ":' _, .. ··- ' ,; ...... ' .. 

.... , 

-

-

•. 

' 

.•·. 

., 

'.
(j "' 

,, 

,, Table 1 (.cont1nuod) • 

M(!tabolic J!& ol Compound Avcror.c NU!nbcr of ll1:Jtidine-Po!l1 tivc ncv(!rtont.s per Pia 

Compound /Jetlvity Added per Plnto 

. " 

ClDA-GtlGX 421-32-D - 1 40 15 0 ::J5 29 1

-.f:l 10 30 15 27 27 1

50 38 ·20 22 27 H

' . - 100 30 13 25 25 1

500 37 1G 30 !G

1000 3G 1G 21 39 15

nooo ·19 10 18 23 1

,, + 1 3G 21 41 39 l5

+ 10 34 12 25 30 l-+ 50 ,.;.. 31 14 3G 3G 1<

co + 100 28 20 41 39 li

,. 31 15 37 40 1

+ 1000 3G 12 3·1 

+ 5000 13 13 21 2G 1G

421-32-C - 1 43 12 29 37 l;

10 29 14 29 27 1

50 30 IG 2G 31 1

100 3G 1<1 28 31 l-

sOb 32 10 1<1 32 1-

1000 40 12 21 3G 1'

sooo .. 23 7 21 23 1

.. 

+ 1 40 15 4G 1G 1

+ 10 3G 10 3G 4t 1-

+ 50 30 10 39 39 l3

+ 100 30 18 43 3G 15

+' 500 33 IS 39 3G l::

,-:_:;_ + 1000 43 18 38 •13 13

+ 5000. IG 15 23 30 13

,. 
•.. ) 

compound 

Cl ll/.•OEI OY MA-26 

"' 

A :3' 

..... -:• C") 0" 

. g: r.f· ;r a--

- ::0 3 I!J ~ ~ o·~ 

"''!.. .u.. !.?.,_g 

.-.. 

iJ 

Mctoba11c .. -~: Aetivlty 

·-

+ 

+ 

+ 

+ 

+ 

+ 

+ 

" -~ Q ~:.: . Tnb1o 1 (Concludedi • 1-lC of Compound J\dded per Plnto •: _. l 10 so 100 500 . 1000 5000 1 10 c~ -- 50 100 500 1000 5000 " - Avcroce Number of lltst1dlne-Pos1ttve Revertants ~ ~·r- -~ ~ 19 9 19 10 31 11 25 43 23 12 • 38 54 18 8 30 11 18 13 25 52 23 11 33 37 39 8 41 33 19 11 23 't~- 36 11 a· 31 3~ 21 15 39 14 14 ·.'~t4 30 17 14 iii 36 11 16 18 31 15 19 ·n 12 15 " ,, " " " " J)cr Pl

t.\-

1

1

1

1

1

1

1G

!5

1

1

1

!5

!5

1<
,. 

• .. 

m:n:m·ll cES 

., 1. J. McC:-mn, E. Cho11 E. Ynr:msak1 1 and B. N. Ames. Detection of 

carcinoccns as mut~~cns in the Salmonclln microsome tCst: Assay of 

300 chcm1c~ls. Proc~. Jlnt. Acad. Sc1. USA ~, 5135-5139. 

• ... 2. B. N .. AmcsJ E. G .. Gurney, J .. A. Miller, and H. Dartsch. Carcinogens :: 

as frarncsh1:Ct mutagens: Mctabol1 tcs and d<!-riv3t1vcs of 2-acctylar.linoflttorcne and other aromatic amine cax·cinogcns. Proc. Nat. Acad. Sc1. 

USA~ 3128-3]32 (1972). " 

3. B. N • .Ames" F. "D. Lc!!, an~ w. E. Dur-ston. An improved b;:,cter1~1 

test system :Cor the detection and cl~ss~!1cat1on of rnuta~ens and 

carcinogens. Proc. Nat. Acad. Sc1. USA 70, 782-786 (1973). 

4. B. N. Ames, E. 

aJ·c mu tagcns: 

activation and 

~ (1973). 

:() 

W. Durston, E. Yamasaki., and F. D • .,Lee. Carcinogens 

A simple test system comb1n1ng liver homogenates for 

bacteria for de'te:ct1on. Pro c. Nat .. .Aoz::ad .. Sci.. USA 

5. J. McCann, N. E. Splng01:r., J .. Kobox:.11 and B. N. Ames. The detection 

o! corcino~ens as ~Ut3ecns: Bacter1~l tester strains with F £actor 

p1asm1ds. P7oc. N~t. Acod. Sc1. US' (1n press, 1975). 

6~ · L. D ... Kicr, E. Ya:r..o.saki 1 ::md B ... N. Ames. 

activity in cigarette smoke condens3tes .. 

71, 4159-4163 (1974) •. 

De tcction of mutagenic. 

Proc. Nat. Acad. 5<:1., USA 

· 7. -;B. N .. Ames, .;r. McCann., omd E. Y01rnasak1.. l •. 1ethods for ' Detecting 

Carcinogens. and Mutagens wl th the Salr.~onella/M::~li'l.mal1:ln-M1crosome 

Mutngen1c1ty Test. J,lut. Res. 317 347-364 (1975)0 

•• 

... 

. :; 

10 

{1·-- .'····~· 

·' 
... 

c:r: 

z 

-.... 

LLI 

~ en 

LLI 

... 

= ... 

; 

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